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1、LETTERdoi:10.1038/nature10693VifhijacksCBF-btodegradeAPOBEC3GandpromoteHIV-1infection1,2344434StefanieJa¨ger*,DongYoungKim*,JuddF.Hultquist*,KeisukeShindo,RebeccaS.LaRue,EunjuKwon,MingLi,4331,2,31,21,22,5BrettD.Anderson,LindaYen,DavidStanley,CathalMahon,JoshuaKa
2、ne,KathyFranks-Skiba,PeterCimermancic,2,32,3,5,62,342,3,61,2,6,7AlmaBurlingame,AndrejSali,CharlesS.Craik,ReubenS.Harris,JohnD.Gross&NevanJ.Krogan13Restrictionfactors,suchastheretroviralcomplementaryDNA(ELOBC)hasbeenreported,thearchitectureofthefull-lengthVifinde
3、aminaseAPOBEC3G,arecellularproteinsthatdominantlyblockcomplexwithhostfactorshasremainedelusive,inpartbecauseVif1–3virusreplication.TheAIDSvirus,humanimmunodeficiencyviruscomplexeshavepoorsolubilityandactivity.Wethereforereasonedtype1(HIV-1),producestheaccessoryf
4、actorVif,whichcounteractsthatVifmaybindanadditionalhostfactorandthatsuchafactormaythehost’santiviraldefencebyhijackingaubiquitinligasecomplex,renderitmoretractableinvitro.containingCUL5,ELOC,ELOBandaRING-boxprotein,andWetookanunbiasedproteomicapproachtoidentifyh
5、ostfactors4–10targetingAPOBEC3Gfordegradation.Herewereveal,usingthatbindall18HIVprocessedandpolyproteinsusinganaffinitytag/anaffinitytag/purificationmassspectrometryapproach,thatVif14,15purificationmassspectrometry(AP–MS)approach.Tothisend,additionallyrecruitsth
6、etranscriptioncofactorCBF-btothis23Strepand33Flagwasfusedtothecarboxy(C)terminusoftheseubiquitinligasecomplex.CBF-b,whichnormallyfunctionsinfactors,includingVif.ThetaggedVifconstructwasbothtransientlyconcertwithRUNXDNAbindingproteins,allowsthereconstitu-transfec
7、tedintoHEK293cellsandusedtomakeastable,tetracycline-tionofarecombinantsix-proteinassemblythatelicitsspecificinducibleVif–Strep–FlagJurkatTcellline(Fig.1a).Epitope-taggedpolyubiquitinationactivitywithAPOBEC3G,butnottherelatedVifwaspurifiedfrombothcelltypesusingan
8、tibodiesspecifictoeitherdeaminaseAPOBEC3A.UsingRNAknockdownandgeneticcom-StreporFlagandaliquotsoftheco-purifyingproteinsweresubjectedplementationstudies,wealsodemonst
